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Adipose Derived Mesenchymal Stem Cell Characteristics in Anal Fistulas

Not Applicable
Completed
Conditions
Adipose Tissue
Perianal Fistula
Tissue Transplantation
Interventions
Procedure: Injection of autologous adipose tissue in anal fistula
Registration Number
NCT04834609
Lead Sponsor
University of Aarhus
Brief Summary

This study investigated the cellular and molecular characteristics of AT-MSCs obtained from autologous AT therapy in patients with high transphincteric perianal fistulas of crytoglandular origin. Adipose tissue was injected into anal fistulas. Characteristics of adipose tissue mesenchymal stemcells (AT-MSC) was investigated and compared in patients with fistula that healed after the treatment (responders) to patients who failed to heal (non-responders)

Detailed Description

Injection with allogene or autologous stem cells has been reported to be efficient treatment of perianal fistulas. An alternative to this treatment could be injection with freshly collected autologous adipose tissue. In this study 27 patients with cryptoglandular anal fistulas were treated with freshly collected autologous adipose tissue.A clinical assessment of the patient prior to inclusion was undertaken and a loose seton placed for at least 6 weeks prior to fat injection. An MRI of the pelvis was performed before inclusion. Fistulas with secondary tracts and/or cavities were excluded. The operation was performed in one procedure including liposuction and injection of adipose tissue. A sample of adipose tissue from all 27 patients was analyzed. AT-MSCs were isolated and characterized using cellular and molecular analyses. Clinical and MRI-scanning evaluation of fistula healing and evaluation of ano-rectal function was performed after 6 months. AT-MSCs phenotype was compared between responders and non-responders with respect to fistula healing. The evaluation of the AT-MSCs was performed in a blinded manner.

Recruitment & Eligibility

Status
COMPLETED
Sex
All
Target Recruitment
27
Inclusion Criteria
  • high trans-sphincteric fistulas
  • fistula confirmed and classified by an MRI.
  • seton (> 6 weeks) prior to fat injection
  • informed, written consent.
Exclusion Criteria

Anovaginal fistula

  • Active sepsis
  • IBD, immunodeficiency, prior pelvic irradiation and malignancy
  • Insulin dependent diabetes
  • More than 4 prior attempts of fistula closure
  • Tobacco smoking or nicotine substitution 8 weeks prior to fat injection.
  • Pregnancy
  • Psychiatric disorders
  • BMI ≥ 35 or BMI<20
  • Active tuberculosis
  • Patient less than 18 years
  • Unable to undergo MRI

Study & Design

Study Type
INTERVENTIONAL
Study Design
SINGLE_GROUP
Arm && Interventions
GroupInterventionDescription
Injection with adipose tissueInjection of autologous adipose tissue in anal fistulaInjection of freshly collected autologous adipose tissue
Primary Outcome Measures
NameTimeMethod
Investigation of differentiation potential of AT-MSCs to differentiate into adipocyteAt start of treatment

Differentiation potential of AT-MSCs: to differentiate into adipocyte measured by Oil-Red O staining and gene expression of adipogenic markers (PPARg and LPL normalized to housekeeping gene beta actin) presented as a Fold change to undifferentiated cells (arbitrary units)

Investigation of differentiation potential of AT-MSCs to differentiate into osteoblastAt start of treatment

Differentiation potential of AT-MSCs: to differentiate into osteoblast measured by Alizarin S staining and gene expression of osteogenic markers (BGALP and RUNX2 normalized to housekeeping gene beta actin) presented as a Fold change to undifferentiated cells (arbitrary units)

Investigation of cell proliferation of AT-MSCsAt start of treatment

Cell proliferation of AT-MSCs evaluated as number of cells/per day

Measurement of gene expression profile of AT-MSCsAt start of treatment

Gene expression of proinflammatory (NFKB, TNFa, IL1B, IL6) and senescence associated molecules(CDKN2A, TP53, TGFB1, VEGFA, IFNG, IL6) of AT-MSCs in relation to the outcome of fistula treatment (i.e. comparison between responders and non-responders). The data are normalized to housekeeping gene beta actin (arbitrary units)

Secondary Outcome Measures
NameTimeMethod
Evaluation of fistula healing after treatment6 months after last injection of autologous adipose tissue

A combination of Clinical and MRI healing defined as closure of the internal and external fistula opening and no discharge and no fluid filled fistula tracts on evaluated as success rate of the healing in (%)

Functional gastroenterological outcome after treatment6 months after last injection of autologous adipose tissue

Anal continence evaluated as the St. Mark's Score (0-24)

Functional urological outcome after treatment6 months after last injection of autologous adipose tissue

Urinary incontinence evaluated as ICIQ-UI-SF (0-21)

Healing of anal fistula after treatment6 months after last injection of autologous adipose tissue

Clinical healing defined as closure of the internal and external fistula opening and no discharge evaluated as success rate of the healing in (%)

Defecation disorder evaluation after treatment6 months after last injection of autologous adipose tissue

Defecation disorders evaluated as Altomare Obstructed Defecation Score (0-31)

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